Neural transplantation in the injured central nervous system (CNS) has had limited success. It is hypothesized that this hostile environment may require improved cell support through directed extracellular matrix (ECM) protein engineering to improve graft survival and cell function and hence, functional recovery. The overall objective of this research project is to develop minimally-invasive transplantation techniques for optimizing stem/progenitor cell attachment through ECM-based scaffolds and to utilize relevant experimental models (both in vitro and in vivo) for optimization and outcome assessment. This overall goal is divided into 3 interrelated specific aims: (1) To characterize neural stem (NS) cell-ECM-based 3-D constructs in vitro for minimally invasive grafting strategies and maximal cell survival and to elicit a desired degree of proliferation, migration, and differentiation; (2) To determine mechanisms of construct integration by testing NS-ECM constructs in a surrogate hostile in vitro environment; and (3) To analyze the in vivo function of tissue-engineered constructs by transplanting constructs into contused mouse brains and examining post-injury alterations in the host contusion, cell behavior, and cognitive and sensorimotor behavioral outcome. The research proposed is significant because it offers a novel approach to progenitor/stem cell transplant technology with detailed analyses of outcome. This research may have direct application to clinical practice in neurosurgery that would permit therapeutic, cellular replacement in the treatment of traumatic brain and spinal cord injuries and degenerative diseases of the CNS. In addition, this research will provide insight into the mechanisms of CNS regeneration and help to elucidate the necessary cellular environment for neurotransplantation success. By analyzing outcome in well-controlled multi-level systems, this research may also lead to acellular transplantation methodology and establish the requirements necessary for the transplantation of non-embryonic/fetal cell sources.